Human BM-MSCs were trans-differentiated into hepatocyte by using two steps protocol: differentiation step followed by maturation step.
FGF8 promotes such enrichment and induces a one-step differentiation toward a unipotential hepatocyte progenitor.
However, such screens have been hampered by heterogeneous neuronal production, high cost and low yield, and multi-step differentiation procedures.
Chen et al. (Chen et al., 2012) established a rapid and efficient three-step differentiation protocol with HGF, activin A, and Wnt3a that is able to generate functional HLCs from human iPSCs.
As expected, CD31 and CD144 were upregulated during the two-step differentiation and expressed robustly after sorting (Figure 2A).
It is known that osteoblasts are derived from mesenchymal stem cells (MSCs) through a multi-step differentiation pathway [46].
Based upon this information, we developed a 4-step differentiation protocol, mimicking four distinct steps of hepatic development (Figure 1).
To increase endothelial differentiation efficiency and avoid contamination from mouse embryonic fibroblasts (MEFs), we developed a novel two-step differentiation process with serum free culture system.
Here we describe a two-step differentiation procedure that takes advantage of collagen I to efficiently promote differentiation of hESCs into endothelial lineage cells.
Our present one-dimensional model cannot describe these priming effects, thus calling for model extensions that account for multi-stable and multi-step differentiation processes in a multidimensional system as suggested in [27].
In summary, our results show that (i) endothelial differentiation efficiency was increased by a two-step differentiation of hESCs and with animal product free procedure by using serum free culture medium, (ii) hESC-ECs possess functional vasculogenic ability in vivo, and (iii) transplantation of hESC-ECs can significantly improve short-term cardiac function at 2 weeks.
